Assessment of functional effect of missense variants. (A,B) Growth assays on solid media (A) and liquid media (B) for human tyrosyl-tRNA synthetase (YARS) gene variants. G41R and E196K are disease-associated variants; M431L and A339T are non-disease-associated variants. The yeast cells were temperature-sensitive mutants of the yeast TYS1 gene, expressing either wild-type or variant alleles of the YARS gene, or the GFP gene as a control. For solid growth assays, fivefold serial dilutions of yeast cells were spotted onto plates and incubated at 24°C and 36°C for 2 d. For liquid growth assays, approximately 105 cells were inoculated in 100 μL liquid medium and the absorbance at 600 nm was read every 15 min. (C) Distribution of solid growth (FCS) scores for disease- and non-disease-associated variants. (D) Distribution of liquid growth (FCT) scores for disease- and non-disease-associated variants. (E) Precision and recall analysis for FCS and PolyPhen2 scores. (F) Overlap of disease-associated variants predicted to be deleterious by functional assays (orange) and PolyPhen2 (green). (G) Overlap of non-disease-associated variants predicted to be deleterious by functional assays (orange) and PolyPhen2 (green). As described in the text, analyses in this figure exclude FCT assays of PKLR variants and all assays of UROS variants.
